Department of Ultrasound, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310058, China.
CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
Signal Transduct Target Ther. 2023 May 8;8(1):182. doi: 10.1038/s41392-023-01398-4.
Gene delivery is the process by which foreign DNA is transferred to host cells, released from intracellular vesicles, and transported to the nuclei for transcription. This process is frequently inefficient and difficult to control spatiotemporally. We developed a gene delivery strategy that uses ultrasound to directly deliver plasmid DNA into nuclei via gas vesicles (GVs)-based intracellular cavitation. pDNA-binding GVs can be taken up by cells and cause intracellular cavitation when exposed to acoustic irradiation and delivering their pDNA payloads into nuclei. Importantly, GVs can remain stable in the cytoplasm in the absence of acoustic irradiation, allowing for temporally controlled nuclear gene delivery. We were able to achieve spatiotemporal control of E-cadherin nuclear gene delivery in this manner, demonstrating its efficacy in tumor invasion and metastasis inhibition. Interestingly, we discovered that nuclear gene delivery of E-cadherin during the G2/M phase of the cell cycle in C6 tumor cells inhibited tumor invasion and metastasis more effectively than during the G1 and S phases. The gene delivery of E-cadherin at the G2/M phase resulted in significantly lower expression of Fam50a, which reduced Fam50a/Runx2 interaction and led to reduced transactivation of MMP13, an important factor for epithelial-mesenchymal transition, as observed in a molecular mechanism assay. Thus, using remote acoustic control of intracellular cavitation of pDNA-GVs, we developed a high spatiotemporally controllable gene delivery strategy and achieved stronger tumor invasion and metastasis inhibition effects by delivering the E-cadherin gene at the G2/M phase.
基因传递是指将外源 DNA 转移到宿主细胞中，从细胞内囊泡中释放出来，并运输到细胞核中进行转录的过程。这个过程通常效率低下，难以进行时空控制。我们开发了一种基因传递策略，该策略使用超声通过基于气穴（GVs）的细胞内空化直接将质粒 DNA 传递到细胞核中。与 pDNA 结合的 GVs 可以被细胞摄取，并在暴露于声辐射时引起细胞内空化，将其 pDNA 有效载荷传递到细胞核中。重要的是，在没有声辐射的情况下，GVs 可以在细胞质中保持稳定，从而实现时间控制的核基因传递。我们能够以这种方式实现 E-cadherin 核基因传递的时空控制，证明其在肿瘤侵袭和转移抑制中的功效。有趣的是，我们发现 C6 肿瘤细胞中 E-cadherin 的核基因传递在细胞周期的 G2/M 期比在 G1 和 S 期更有效地抑制肿瘤侵袭和转移。在 G2/M 期进行 E-cadherin 基因传递会导致 Fam50a 的表达显著降低，这降低了 Fam50a/Runx2 相互作用，并导致 MMP13 的转录激活减少，MMP13 是上皮-间充质转化的一个重要因素，如分子机制分析中观察到的那样。因此，我们通过远程声控 pDNA-GVs 的细胞内空化，开发了一种高时空可控的基因传递策略，并通过在 G2/M 期传递 E-cadherin 基因实现了更强的肿瘤侵袭和转移抑制效果。
